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International audience; Nitrous acid (HONO) photolysis is an important atmospheric reaction that leads to the formation of hydroxyl radicals (OH), the main diurnal atmospheric oxidants. The process of HONO formation remains unclear, and comparisons between field measurements and model results have highlighted the presence of unknown HONO sources. HONO production on plant surfaces was recently suggested to contribute to atmospheric HONO formation, but there is limited information on the quantification of HONO production and uptake by plants. To address this gap in the existing knowledge, the current study investigated HONO exchange on living Zea mays plants. Experiments were conducted in growth chambers under controlled experimental conditions (temperature, relative humidity, NO2 mixing ratio, light intensity, CO2 mixing ratio) at temperatures ranging between 283 and 299 K. To investigate the effect of drought on HONO plant-atmosphere exchanges, experiments were carried out on two sets of Zea mays plants exposed to two different water supply conditions during their growth: optimal watering (70% of the field capacity) and water stress (30% of the field capacity). Results indicated that the uptake of HONO by control Zea mays plants increased linearly with ambient temperature, and was correlated with CO2 assimilation for temperatures ranging from 283 to 299 K. At 299 K, HONO production on the leaves offset this uptake and Zea mays plants were a source of HONO, with a net production rate of 27 ± 7 ppt h-1. Deposition velocities were higher for HONO than CO2, suggesting a higher mesophyll resistance for CO2 than HONO. As water stress reduced the stomatal opening, it also decreased plant-atmosphere gas exchange. Thus, climate change, which may limit the availability of water, will have an impact on HONO exchange between plants and the atmosphere.

International audience; Soil microorganisms play a key role in soil organic matter (SOM) dynamics, but little is known about the controls affecting the distribution of microbial biomass and their residues in soil. Here, a forested Cambisol topsoil was incubated with C-13-labeled glycine or beech leaves for 12 weeks prior to sequential density fractionation. The incorporation of the C-13 label in amino sugars (AS) was used to gain insight into bacterial and fungal assimilation of the substrates. AS derived from glycine or leaves were compared to total AS to investigate how microbial residues and active communities were distributed among soil density fractions.Bacteria slightly dominated leaf C assimilation, while a pronounced fungal dominance was observed for glycine. The glycine-derived AS and original AS were similarly distributed among the soil density fractions, both peaking in microbial aggregates (1.8-2.4 g cm(-3)). Leaf-derived AS were mostly found in association with the plant debris (

13 pages; International audience; The turnover of organic matter in soil depends on the activity of microbial decomposers. However, little is known about how modifications of the diversity of soil microbial communities induced by fresh organic matter (FOM) inputs can regulate carbon cycling. Here, we investigated the decomposition of two 13C labeled crop residues (wheat and alfalfa) and the dynamics of the genetic structure and taxonomic composition of the soil bacterial communities decomposing 13C labeled FOM and native unlabeled soil organic matter (SOM), respectively. It was achieved by combining the stable isotope probing method with molecular tools (DNA genotyping and pyrosequencing of 16S rDNA). Although a priming effect (PE) was always induced by residue addition, its intensity increased with the degradability of the plant residue. The input of both wheat and alfalfa residues induced a rapid dynamics of FOM-degrading communities, corresponding to the stimulation of bacterial phyla which have been previously described as copiotrophic organisms. However, the dynamics and the identity of the bacterial groups stimulated depended on the residue added, with Firmicutes dominating in the wheat treatment and Proteobacteria dominating in the alfalfa treatment after 3 days of incubation. In both treatments, SOMdegrading communities were dominated by Acidobacteria, Verrucomicrobia, and Gemmatimonadetes phyla which have been previously described as oligotrophic organisms. An early stimulation of SOM-degrading populations mainly belonging to Firmicutes and Bacteroidetes groups was observed in the alfalfa treatment whereas no change occurred in the wheat treatment. Our findings support the hypothesis that the succession of bacterial taxonomic groups occurring in SOM- and FOMdegrading communities during the degradation process may be an important driver of the PE, and consequently of carbon dynamics in soil.

Enzymes of the DMSO reductase family use a mononuclear Mo-bis(molybdopterin) cofactor (MoCo) to catalyze a variety of oxo-transfer reactions. Much functional information on nitrate reductase, one of the most studied members of this family, has been gained from EPR spectroscopy, but this technique is not always conclusive because the signature of the MoCo is heterogeneous, and which signals correspond to active species is still unsure. We used site-directed mutagenesis, EPR and protein film voltammetry to demonstrate that the MoCo in R. sphaeroides periplasmic nitrate reductase (NapAB) is subject to an irreversible reductive activation process whose kinetics we precisely define. This activation quantitatively correlates with the disappearance of the so-called "Mo(V) high-g" EPR signal, but this reductive process is too slow to be part of the normal catalytic cycle. Therefore, in NapAB, this most intense and most commonly observed signature of the MoCo arises from a dead-end, inactive state that gives a catalytically competent species only after reduction. This activation proceeds, even without substrate, according to a reduction followed by an irreversible nonredox step, both of which are pH independent. An apparently similar process occurs in other nitrate reductases (both assimilatory and membrane bound), and this also recalls the redox cycling procedure, which activates periplasmic DMSO reductases and simplifies their spectroscopic signatures. Hence we propose that heterogeneity at the active site and reductive activation are common properties of enzymes from the DMSO reductase family. Regarding NapAB, the fact that we could detect no Mo EPR signal upon reoxidizing the fully reduced enzyme suggests that the catalytically active form of the Mo(V) is thermodynamically unstable, as is the case for other enzymes of the DMSO reductase family. Our original approach, which combines spectroscopy and protein film voltammetry, proves useful for discriminating the forms of the active site on the basis of their catalytic properties. This could be applied to other enzymes for which the question arises as to the catalytic relevance of certain long-lived, spectroscopically characterized species.

International audience

Background YedY, a molybdoenzyme belonging to the sulfite oxidase family, is found in most Gram-negative bacteria. It contains a twin-arginine signal sequence that is cleaved after its translocation into the periplasm. Despite a weak reductase activity with substrates such as dimethyl sulfoxide or trimethylamine N-oxide, its natural substrate and its role in the cell remain unknown. Although sequence conservation of the YedY family displays a strictly conserved hydrophobic C-terminal residue, all known studies on Escherichia coli YedY have been performed with an enzyme containing a 6 histidine-tag at the C-terminus which could hamper enzyme activity. Results In this study, we demonstrate that the tag fused to the C-terminus of Rhodobacter sphaeroides YedY is detrimental to the enzyme’s reductase activity and results in an eight-fold decrease in catalytic efficiency. Nonetheless this C-terminal tag does not influence the properties of the molybdenum active site, as assayed by EPR spectroscopy. When a cleavable His-tag was fused to the N-terminus of the mature enzyme in the absence of the signal sequence, YedY was expressed and folded with its cofactor. However, when the signal sequence was added upstream of the N-ter tag, the amount of enzyme produced was approximately ten-fold higher. Conclusion Our study thus underscores the risk of using a C-terminus tagged enzyme while studying YedY, and presents an alternative strategy to express signal sequence-containing enzymes with an N-terminal tag. It brings new insights into molybdoenzyme maturation in R. sphaeroides showing that for some enzymes, maturation can occur in the absence of the signal sequence but that its presence is required for high expression of active enzyme.